Supplementary Methods, Figures 1 - 4, Tables 2 - 6 from The Noncoding RNA <i>MALAT1</i> Is a Critical Regulator of the Metastasis Phenotype of Lung Cancer Cells

PDF file - 808K, Supplementary Figure S1: This figure depicts the impact of MALAT1 loss on gene expression, splicing and SR protein phosphorylation. Supplementary Figure S2: This figure describes the generation of MALAT1-Knockout A549 cells using an independent ZFN as well as their phenotypes regarding proliferation and MALAT1 target gene expression. Supplementary Figure S3: This figure displays the impact of loss of MALAT1 using an independent ZFN in A549 cells on cell migration. Supplementary Figure S4: This figure documents that Antisense Oligonucleotides (ASOs) are effective in silencing MALAT1 in vitro without affecting cell proliferation of EBC-1 cells. Supplementary Table S2: This table lists 23 genes whose differential expression (5 up / 18 down) in MALAT1 KO cells was validated via qRT-PCR and who had been previously identified with critical roles in lung cancer, migration, invasion, EMT or metastasis. Supplementary Table S3: This table lists the sequences of all primers used for qRT-PCR in 5'-3' direction. Supplementary Table S4: This table lists the sequences of all primers used for splicing analysis in 5'-3' direction. Supplementary Table S5: This table lists the sequences of all primers and probes used for TaqMan qRT-PCR in 5'-3' direction. Supplementary Table S6: This table lists the sequences of all Antisense Oligonucleotides (ASOs) in 5'-3' direction. Supplementary Methods: This chapter provides experimental details on the RNA isolation and DNase I digest, reverse transcription and quantitative PCR, exon microarray analysis, antisense oligonucleotides (ASOs), lung imaging by microCT scan, immunohistochemistry and in situ hybridization, statistical analysis, generation of ZFN-2 clones, cell proliferation assays, SDS-PAGE and Western blotting.